CN110903367B - Application of transcription factor in increasing opening angle of apple branches - Google Patents
Application of transcription factor in increasing opening angle of apple branches Download PDFInfo
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- 108091023040 Transcription factor Proteins 0.000 title claims abstract description 14
- 102000040945 Transcription factor Human genes 0.000 title claims abstract description 14
- 235000011430 Malus pumila Nutrition 0.000 claims description 17
- 235000015103 Malus silvestris Nutrition 0.000 claims description 17
- 239000002773 nucleotide Substances 0.000 claims description 6
- 125000003729 nucleotide group Chemical group 0.000 claims description 6
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 4
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 2
- 244000141359 Malus pumila Species 0.000 claims description 2
- 241000220225 Malus Species 0.000 abstract description 24
- 230000002018 overexpression Effects 0.000 abstract description 13
- 235000021016 apples Nutrition 0.000 abstract description 8
- 241000196324 Embryophyta Species 0.000 abstract description 6
- 238000010353 genetic engineering Methods 0.000 abstract description 2
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- 238000000034 method Methods 0.000 description 2
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- 206010067484 Adverse reaction Diseases 0.000 description 1
- 101100132373 Arabidopsis thaliana MYB88 gene Proteins 0.000 description 1
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- 235000004789 Rosa xanthina Nutrition 0.000 description 1
- 241000220222 Rosaceae Species 0.000 description 1
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- 238000009825 accumulation Methods 0.000 description 1
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- 150000001413 amino acids Chemical class 0.000 description 1
- 229930014669 anthocyanidin Natural products 0.000 description 1
- 150000001452 anthocyanidin derivatives Chemical class 0.000 description 1
- 235000008758 anthocyanidins Nutrition 0.000 description 1
- 239000002363 auxin Substances 0.000 description 1
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- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
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Abstract
The invention belongs to the technical field of biotechnology and plant genetic engineering, and discloses application of a transcription factor in increasing the apple branch opening angle. When measured by a protractor, the average angle of the branches of the Gala apples without transgenes is 54.25 degrees, the average opening angle of the branches of the MdMYB124 overexpression strains #1, #2, #19 is 79 degrees, 74.9 degrees and 81.8 degrees, and the average opening angle of the overexpression strain #23 of the MdMYB88 is 83.6 degrees.
Description
Technical Field
The invention belongs to the technical field of biotechnology and plant genetic engineering, and particularly relates to application of a transcription factor in increasing the opening angle of apple branches.
Background
Currently, the closest prior art: the plant type with large opening angle can reduce the mutual shielding among the leaves, so that the leaves can uniformly receive illumination, thereby improving the photosynthetic efficiency of the leaves and increasing the carbon content of the leaves. The correlation and regression analysis results of the content of the nutrient substances in the leaves of the fruit trees and the indexes of the fruit quality show that the content of the carbon nutrient substances in the leaves after the branches of the fruit trees are treated by different branch pulling angles is positively correlated with the weight of a single fruit, the fruit shape index, the anthocyanidin content, the hardness, the vC content, the total sugar content and the protein content of the fruit and is negatively correlated with the total acid content of the fruit. Therefore, the fruit quality can be improved by improving the opening angle of branches and leaves. The rape branch angle finds 27 genes as candidate genes through QTL, including auxin response genes, and provides basis for deciphering the genetic control of the rape branch angle. After artificial branch pulling treatment, branch angles of branches become large, the growth direction of the branches is changed, the GA and IAA content on the upper sides of the branches is obviously increased, the GA and IAA content on the lower sides of the branches is obviously reduced, and the GA and IAA content on the upper sides of the same branches is obviously higher than that on the lower sides of the same branches, so that the GA and the IAA have important influence on the branch angles of the branches in the branch growing and developing process.
In summary, the problems of the prior art are as follows: in the prior art, the cost is increased through manual work, medicament, material management and the like, and the technical implementation difficulty is high.
The difficulty of solving the technical problems is as follows: the hormone has double-side effect, and small dosage can not play a role, while large dosage can play a role in adverse reaction. The branch pulling time and the branch pulling angle are required to be carried out by professional fruit tree practitioners. MYB88 transgenic material provides scion for cultivating apple varieties with large opening angles or hybrid parents. And provides experimental materials for researching the increase of the opening angle of the apple.
The significance of solving the technical problems is as follows: the invention simplifies the management and reduces the cost.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a transcription factor, an expression vector and application in increasing the fruit tree branch opening angle.
The invention is realized in such a way that the transcription factor for increasing the fruit tree branch opening angle is characterized in that the transcription factor MdMYB88/124 for increasing the fruit tree branch opening angle has the sequence: SEQ ID NO: 1-SEQ ID NO: 4.
the invention also aims to provide an expression vector constructed by the transcription factor for increasing the fruit tree branch opening angle, wherein the expression vector comprises the following components in parts by weight: pCAMBIA2300, pGWB414, pGWB 415.
The invention also aims to provide an application of the transcription factor in increasing the fruit tree branch opening angle.
In summary, the advantages and positive effects of the invention are: according to the invention, an important transcription factor MdMYB88/124 in the apples is discovered through a transgenic means, after overexpression, the branch opening angle of the apples is obviously increased, the fruit tree opening angle is increased, the significance for fruit tree production is great, ventilation and light transmission are increased, the yield is improved, manual branch pulling is replaced, and the orchard management cost is saved. When the transgenic Gala apple branches are measured by a protractor, the average angle of the transgenic Gala apple branches is 64 degrees, the average opening angle of the opened branches of the MdMYB124 overexpression strain # 1 is 85 degrees, the average opening angle of the opened branches of the MdMYB124 overexpression strains # 1, #18 and #19 is 85 degrees, 80 degrees and 82 degrees, the average opening angle of the overexpression strain # 23 of the MdMYB88 is 70 degrees, and the included angles of the branches are obviously increased by overexpression. Silencing lines # 24, #25 significantly reduced shoot angles of 54 degrees and 44 degrees, respectively.
And the included angle of the leaves is measured, MdMYB88 not only has the function of increasing the included angle of branches, but also can increase the included angle of the leaves of the apples. The included angle formed by the petiole and the stem of the branch is measured, the transgenic gala is absent, the included angle of the leaf is 55 degrees, the over-expressed strain is #29, and the average included angle of the #10 leaf is 68 degrees and 63 degrees, so that the included angle of the gala leaf is obviously increased. The leaf angle of the silent strain # 24, #25 was significantly reduced by an average of 44 degrees and 46 degrees. The included angle of the petioles becomes larger.
According to the invention, by increasing the mechanism that fruit bearing is influenced by the opening angle of the branches of the fruit trees, the study in the fifth and sixth decades of the 20 th century is clear, the branch pulling of the young trees can promote the crown expansion and early formation, the branch pulling of the large trees can improve the ventilation and light transmission conditions of the tree bodies, promote the illumination of the inner cavities of the fruit trees, and improve the light energy utilization rate, thereby promoting the fruit coloring and improving the yield. Recent studies have found that external application of phytohormones can also increase the included angle of plant branches. MdMYB88 is a gene playing an important role in regulating and controlling the growth and development of plants and the stress of abiotic stress. When MdMYB88 transgenic apple material is observed, the fact that the leaf included angle of seedlings and the opening angle of branches growing in two years of an over-expression strain MdMYB88 are both large is significant for fruit tree production, ventilation and light transmission are increased, the light transmission area is increased, the overall photosynthetic rate is improved, nutrition accumulation is promoted, the yield is improved, the like, manual branch pulling is replaced, fruit tree pruning is reduced, and orchard management cost is saved.
According to the invention, the function of increasing the opening angle of branches and leaves of the MdMYB88 transgenic strain is researched, so that the method plays an important role in actual production and new variety cultivation.
Drawings
FIG. 1 is a schematic diagram of an included angle between a Gala apple and a transgenic branch according to an embodiment of the present invention;
in the figure: (a) GL-3; (b) MdMYB124 OE # 19; (c) MdMYB124 OE # 18; (d) MdMYB124 OE # 1; (e) MdMYB88 OE # 23; (f) MdMYB88/124RNAi #25(g) MdMYB88/124RNAi # 24
FIG. 2 is a histogram of the included angle between the Gala apple and the transgenic branch provided by the embodiment of the present invention.
FIG. 3 is a schematic view of the included angle of the petioles provided by an embodiment of the present invention;
FIG. 4 is a schematic leaf angle of MdMYB88 transgenic plants and GL-3 provided by an embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The following detailed description of the principles of the invention is provided in connection with the accompanying drawings.
The sequence of the transcription factor provided by the embodiment of the invention is as follows: the nucleotide sequence of MdMYB88 is SEQ ID NO: 1; the amino acid sequence of MdMYB88 is SEQ ID NO: 2; the nucleotide sequence of MdMYB124 is SEQ ID NO: 3; the nucleotide sequence of MdMYB124 is SEQ ID NO: 4. the amino acid sequence of MdMYB88 is SEQ ID NO: 2MdMYB88(XP _008389496.1) and MdMYB124(XP _ 008339925.1). According to apple genome structure annotations provided by a Rosaceae genome website (https:// www.rosaceae.org/; GDR), the full lengths of genes MdMYB88(MDP0000165523) and MdMYB124(MDP0000210970) of apples are 1620bp and 1572bp respectively, while in the apple gene structure annotations provided by an NCBI (https:// www.ncbi.nlm.nih.gov /) database, the lengths of MdMYB88(XP _008389496.1) and MdMYB124(XP _008339925.1) are 1413bp and 1410bp respectively, the nucleotide similarity of the genes MdMYB88 and MdMYB124 is up to 91.49%, and the amino acid similarity is up to 87.95%.
The following describes the effects of the present invention in detail with reference to the accompanying drawings.
MdMYB88/124 is a homologous gene in apple, and has redundant functions. Genetic transformation was performed in the Royal Gala by leaf disc transformation to obtain transgenic seedlings, overexpression lines # 23, #29 of MdMYB124 overexpression lines # 1, #18, #19 and MdMYB 88. Through observation of comparison between the two-year-old transgenic seedlings and the Royal Gala apples, the fact that when MdMYB88 or MdMYB124 is overexpressed in the Gala apples is found that the opening angle of branches is larger than that of the Gala apple trees. When the transgenic Gala apple branches are measured by an angle measurer, the average angle of the transgenic Gala apple branches is 64 degrees, the average opening angle of the branches of MdMYB124 overexpression strains # 1, #18 and #19 is 85 degrees, 80 degrees and 82 degrees, the average opening angle of the overexpression strain # 23 of MdMYB88 is 70 degrees, and the included angle of the branches is obviously increased by overexpression. Silencing lines # 24, #25 significantly reduced shoot angles of 54 degrees and 44 degrees, respectively.
And the included angle of the leaves is measured, MdMYB88 not only has the function of increasing the included angle of branches, but also can increase the included angle of the leaves of the apples. The included angle formed by the petiole and the stem of the branch is measured, the transgenic gala is absent, the included angle of the leaf is 55 degrees, the over-expressed strain is #29, and the average included angle of the #10 leaf is 68 degrees and 63 degrees, so that the included angle of the gala leaf is obviously increased. The leaf angle of the silent strain # 24, #25 was significantly reduced by an average of 44 degrees and 46 degrees. The included angle of the petioles becomes larger.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
<110> Tuqingmei, Liu Xiaofang, Xieyingpeng, Zhaocaide, Xu\22426, Wang Bian, Zhang \29740Ting, Zhang Danni
Application of <120> transcription factor in increasing opening angle of apple branches
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ATGCCGCAGGAGGAGTCAAAGAAGAAGGAGCGCCATATCGTTACTTGGTCTCAAGAGGAGGATGACATACTGAGGAATCAAATAAGCCTCCATGGAACAGAAAATTGGGCTGTCATTGCATCTAAATTCAAGGATAAAACAACCAGACAGTGCAGGAGAAGATGGTACACATACTTGAATTCTGACTTCAAGAAGGGAGGATGGTCACCAGAGGAAGACATGCTCTTGTGCGAGGCTCAGAAGATATTCGGGAACAGATGGACAGAAATAGCAAAGGTGGTTTCAGGAAGAACTGACAATGCCGTGAAAAACCGGTTTTCAACTTTGTGCAAGAAGAGAGCAAAATATGAGGCCTTAGCTAAAGAAAATGCAAATTCGTACATTAATCAAAATGACAAAAGGGTCATAATACGAAATGGATTCAATACAGATGGAACAACAGAAACAACAGCACCACCTAAGAAGATGAGGAGAAGCCACATCCCTATTCTCTCCAAAAATTGCACCTCTGGGGACAGACTACCTGATCAGTGTGGAAAGATAAATCAGCAGCTAAGACCTCCGTTTACTGTGTTGATTCAGAATGCTCACAATGTAGAGAATCTGCAAGACCAGCATAACGGGCATAGCAYCAAGGAAGTCCCCGGGAATGCAGTACAAAATAGCAAGTTTCAAGGATCATTTCTCGAGAAGGATGACCCGAAGATAATTGCTTTGATGCAACAAGCGGAGTTGCTCAGCTCGCTGGCATTAAAAGTTAACTCAGAGAACACAGACCAGAGTCTTGAAAATGCTTGGATGGTGCTCCAAGATTTCTTGAATCAAAGCAAAGATAGTGACATCCTCAGTTATGGAATTGACGATTTTGATTTTCAACTCGAAGATCTTAAATATCTGTTAGAGGACTTAAGGAGTACTACTGAGGGAAGCCGACCATCTTGGCAGCAGCCAGATCTATTCGACGACTCTCCAGGAAGTTCAGAATACAGTACAGGGTCAACTCTTCTGTCCCAAACAGAATGCTATCAAGTGGAACAAAATGAAGTTGAAATAGGTTCACCGAATCAGGAGATTCGACTAGGGTCACACTCAATTCCTATTGAAGGGCAAAATGATGTTRGTGATTGTGAGAAAGGGATTGTTTCCGAAAAGCAAGAGATATTTCCATCATGTGATGAAGGAAGAAAAGACTACGCAGTTGTTTCTGCGTTGTCAAATACCGAGTTCACTTCTCCTTTTAAAGTTGTCCCATTGTTCAGATCCTTGGCAGCAGGAATTCCTAGCCCAAAATTTTCTGAAAGTGAGAGGAATTTCTTGCTCAAAACGCTTGGAGAGGACTCCCCCTGCCCCATCCCAAGCACCAATCCTTCACAACCGCCACCCTGCAAACGATCCCTCCTCCAAAGTCTATAA
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MPQEESKKKERHIVSWSQEEDDILRNQINTHGTENWAIIASKFKDKTTRQCRRRWYTYLNSDFKKGGWSPEEDMLLCEAQKIFGNRWTEIAKVVSGRTDNAVKNRFSTLCKKRAKYEALAKENAASYINQNDKRVKIRNGFNTDGTAETTAPSKKMRRSHIPSLSEGDRLLEQCGKMSQQLRAPFAVLIQNVHNVENLPDQNNVNGTKEVPVNAVQKSKFHGSFLKKDDPKIIALMQQAELLSSLALKVNAENTDQSLENAWKVLQDFLNQSKDSDILSYGINDFDFQLEDLKYLLEDLRSTTEGSRPSWQHCRQPDLYEESPGSSEYSTGSTLLSQTEYYQVEKNEVEIGSLNQEIRPGSQSIPIEGKNGVGDCEKGIFSEKQEIFPSCDEATKDYAVVSALSSIEFNSPIKVTPLFRSLAAGIPSPKFSESERNFLLKTLGEDSPCPNPSTNPSQPPPCKRSLLQSL
Claims (1)
1. Application of transcription factors MdMYB88 and MdMYB124 in increasing the opening angle of branches of apple trees;
the nucleotide sequence of the transcription factor MdMYB88 is SEQ ID NO: 1, amino acid sequence of SEQ ID NO: 2; the nucleotide sequence of MdMYB124 is SEQ ID NO: 3, the amino acid sequence is SEQ ID NO: 4.
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| CN106086039A (en) * | 2016-08-25 | 2016-11-09 | 上海交通大学 | A kind of Herba Artemisiae Annuae WRKY class transcription factor coded sequence and application |
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| CN108998470B (en) * | 2018-08-06 | 2022-02-11 | 南京农业大学 | Application of soybean MYB32 transcription factor coding gene GmMYB32 |
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|---|---|---|---|---|
| CN106086039A (en) * | 2016-08-25 | 2016-11-09 | 上海交通大学 | A kind of Herba Artemisiae Annuae WRKY class transcription factor coded sequence and application |
| CN108530524A (en) * | 2018-04-18 | 2018-09-14 | 山东省果树研究所 | The application of birch-leaf pear Pb4RMYB genes and its coding albumen in improving plant salt endurance |
Non-Patent Citations (4)
| Title |
|---|
| MdMYB88 and MdMYB124 Enhance Drought Tolerance by Modulating Root Vessels and Cell Walls in Apple;Dali Geng等;《Plant Physiology》;20180906;第178卷(第3期);第1296-1309页 * |
| Ｒ2Ｒ3 型 MYB 转录因子 HbMYB88 结构和功能分析;樊松乐 等;《植物研究》;20191023;第40卷(第1期);第106-116页 * |
| transcription factor MYB51 isoform X1 [Malus domestica];unkown;《GENEBANK DATABASE》;20190503;ORIGIN * |
| transcription factor MYB88 isoform X1 [Malus domestica];unkown;《GENEBANK DATABASE》;20190503;ORIGIN * |
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| CN110903367A (en) | 2020-03-24 |
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